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Dissertations / Theses on the topic 'Immortalized cell line'

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Published: 9 March 2023

Last updated: 10 March 2023

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1

Valtink, Monika, Rita Gruschwitz, Richard H. W. Funk, and Katrin Engelmann. "Two Clonal Cell Lines of Immortalized Human Corneal Endothelial Cells Show either Differentiated or Precursor Cell Characteristics." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-136199.

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Access to primary human corneal endothelial cells (HCEC) is limited and donor-derived differences between cultures exacerbate the issue of data reproducibility, whereas cell lines can provide sufficient numbers of homogenous cells for multiple experiments. An immortalized HCEC population was adapted to serum-free culture medium and repeated cloning was performed. Clonally grown cells were propagated under serum-free conditions and growth curves were recorded. Cells were characterized immunocytochemically for junctional proteins, collagens, Na,K-ATPase and HCEC-specific 9.3.E-antigen. Ultrastructure was monitored by scanning and transmission electron microscopy. Two clonal cell lines, HCEC-B4G12 and HCEC-H9C1, could be isolated and expanded, which differed morphologically: B4G12 cells were polygonal, strongly adherent and formed a strict monolayer, H9C1 cells were less adherent and formed floating spheres. The generation time of B4G12 cells was 62.26 ± 14.5 h and that of H9C1 cells 44.05 ± 5.05 h. Scanning electron microscopy revealed that B4G12 cells had a smooth cell surface, while H9C1 cells had numerous thin filopodia. Both cell lines expressed ZO-1 and occludin adequately, and little but well detectable amounts of connexin-43. Expression of HCEC-specific 9.3.E-antigen was found commensurately in both cell lines, while expression of Na,K-ATPase α1 was higher in H9C1 cells than in B4G12 cells. B4G12 cells expressed collagen IV abundantly and almost no collagen III, while H9C1 cells expressed both collagens at reasonable amounts. It is concluded that the clonal cell line B4G12 represents an ideal model of differentiated HCEC, while H9C1 may reflect features of developing or transitional HCEC
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich

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2

Valtink, Monika, Rita Gruschwitz, Richard H. W. Funk, and Katrin Engelmann. "Two Clonal Cell Lines of Immortalized Human Corneal Endothelial Cells Show either Differentiated or Precursor Cell Characteristics." Karger, 2008. https://tud.qucosa.de/id/qucosa%3A27701.

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Access to primary human corneal endothelial cells (HCEC) is limited and donor-derived differences between cultures exacerbate the issue of data reproducibility, whereas cell lines can provide sufficient numbers of homogenous cells for multiple experiments. An immortalized HCEC population was adapted to serum-free culture medium and repeated cloning was performed. Clonally grown cells were propagated under serum-free conditions and growth curves were recorded. Cells were characterized immunocytochemically for junctional proteins, collagens, Na,K-ATPase and HCEC-specific 9.3.E-antigen. Ultrastructure was monitored by scanning and transmission electron microscopy. Two clonal cell lines, HCEC-B4G12 and HCEC-H9C1, could be isolated and expanded, which differed morphologically: B4G12 cells were polygonal, strongly adherent and formed a strict monolayer, H9C1 cells were less adherent and formed floating spheres. The generation time of B4G12 cells was 62.26 ± 14.5 h and that of H9C1 cells 44.05 ± 5.05 h. Scanning electron microscopy revealed that B4G12 cells had a smooth cell surface, while H9C1 cells had numerous thin filopodia. Both cell lines expressed ZO-1 and occludin adequately, and little but well detectable amounts of connexin-43. Expression of HCEC-specific 9.3.E-antigen was found commensurately in both cell lines, while expression of Na,K-ATPase α1 was higher in H9C1 cells than in B4G12 cells. B4G12 cells expressed collagen IV abundantly and almost no collagen III, while H9C1 cells expressed both collagens at reasonable amounts. It is concluded that the clonal cell line B4G12 represents an ideal model of differentiated HCEC, while H9C1 may reflect features of developing or transitional HCEC.
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

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3

Thieme, Sebastian, Alexander Holzbaur, Ralf Wiedemuth, Aline Binner, Katrin Navratiel, Konstantinos Anastassiadis, Sebastian Brenner, and Cornelia Richter. "The Dox-pDC - A murine conditionally immortalized plasmacytoid dendritic cell line with native immune profile." Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2018. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-234947.

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Plasmacytoid dendritic cells (pDC) constitute a very rare blood cell population and play a significant role in immune response and immune-mediated disorders. Investigations on primary pDCs are hindered not only due to their rarity but also because they represent a heterogeneous cell population which is difficult to culture ex vivo. We generated a conditionally immortalized pDC line (Dox-pDC) from mice with Doxycycline-inducible SV40 Large T Antigen with a comparable immune profile to primary pDCs. The Dox-pDC secrete pro- and anti-inflammatory cytokines upon Toll-like receptor 9 stimulation and upregulate their MHCI, MHCII and costimulatory molecules. Further, the Dox-pDC activate and polarize naïve T cells in vivo and in vitro in response to the model antigen Ovalbumin. Due to their long-term culture stability and their robust proliferation Dox-pDC represent a reliable alternative to primary mouse pDC.

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4

Yin, Zhanhai. "Establishment of a clonal immortalized human mesenchymal stem cell line expressing hTERT using lentiviral gene transfer." Diss., lmu, 2012. http://nbn-resolving.de/urn:nbn:de:bvb:19-145290.

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5

HOSHINO, MUNEMITSU, MUTSUSHI MATSUYAMA, OSAMU TAGUCHI, MORIAKI KUSAKABE, WORAWIDH WAJJWALKU, JIN LU, TOYOHARU YOKOI, et al. "Establishment and Characterization of Immortalized Non-Transplantable Mouse Mammary Cell Lines Cloned from a MMTV-induced Tumor Cell Line Cultured for A Long Duration." Nagoya University School of Medicine, 1991. http://hdl.handle.net/2237/17515.

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6

Wang, Tianren. "The role of SHIP2 in suppressing inflammatory signaling induced by LPS in immortalized murine macrophage cell line." Thesis, University of British Columbia, 2016. http://hdl.handle.net/2429/56632.

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Inflammation is an important step in the body’s defense against pathogen infection. However, it must be tightly regulated and appropriately terminated to prevent pathological consequences. Interleukin-10 (IL10) is one of the body’s most important anti-inflammatory cytokine that can inhibit many molecular events necessary for promoting inflammation including production of pro-inflammatory cytokines such as Tumor Necrosis Factor α (TNFα). Our laboratory has recently shown that SH2-domain containing Inositol 5ʹ phosphatase (SHIP1) is involved in IL10 signaling in macrophages, and although the mechanism of how this occurs is not well studied, our laboratory have obtained data suggesting SHIP1 mediates IL10 signalling through its phosphatase activity or interaction with other signalling proteins. SHIP2 is the only other known homologue of SHIP1 with approximately 38% amino acid sequence identity, yet they possess several similar functions including mediating FcγIIB signaling and phagocytosis. Because of their similarities and SHIP1’s involvement in IL10 signaling, we sought to investigate whether SHIP2 is also involved in inhibiting inflammatory response in macrophage by knocking it out using CRISPR/Cas9-mediated genome editing. Overall, we were unable to determine whether SHIP2 plays a role in macrophage anti-inflammatory response due to the large variation in cell sensitivity to IL10 and we also observed that transduction of macrophages with CRISPR/Cas9 virus alters the cellular response to IL10 which confounded our investigation of SHIP2 function.
Medicine, Faculty of
Biochemistry and Molecular Biology, Department of
Graduate

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7

Rekus, Martin T. "Characterization of growth and differentiation of a spontaneously immortalized keratinocyte cell line (HaCaT) in a defined, serum free culture system." [S.l.] : [s.n.], 2000. http://deposit.ddb.de/cgi-bin/dokserv?idn=963505262.

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8

Yin, Zhanhai [Verfasser], and Wolf [Akademischer Betreuer] Mutschler. "Establishment of a clonal immortalized human mesenchymal stem cell line expressing hTERT using lentiviral gene transfer : no / Zhanhai Yin. Betreuer: Wolf Mutschler." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2012. http://d-nb.info/1024658546/34.

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9

Otsuka, Toshiyuki. "Regulated expression of neurogenic basic helix-loop-helix transcription factors during differentiation of the immortalized neuronal progenitor cell line HC2S2 into neurons." Kyoto University, 1998. http://hdl.handle.net/2433/182245.

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10

CAMBIANICA, ILARIA NADIA. "In vitro blood brain barrier models as a screening tool for brain targeted nanobased drug delivery systems." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2012. http://hdl.handle.net/10281/39834.

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The blood brain barrier (BBB) is a selective biological barrier located at the brain capillaries, that protects the central nervous system (CNS) by monitoring exchanges between blood and brain. The BBB controls and regulates the composition of the CNS environment and it still constitutes the main obstacle for drug delivery to the brain (Weiss N. et al., 2009). The significant scientific and industrial interest in the physiology and pathology of the BBB led to the development of vast number of in vitro BBB models. Even though no “ideal” model exists yet, some of the currently available ones are very useful to investigate permeability, transport mechanisms and cellular and molecular events which occur at the BBB level. New strategies for brain targeted drug delivery exploit endogenously expressed transporters to elicit drug passage across the BBB. Among them, nanoparticles represent a promising tool, since they are biocompatible and biodegradable, and they can be functionalized to target the BBB (De Boer A.G. and Gaillard P.J., 2007) (Beija M et al., 2012; Caruthers S.D. et al., 2007; Moghimi S.M. et al., 2005). In this study we settled in vitro BBB models to identify, with high-throughput screening, the most promising nanoliposomes (NL) for combined BBB crossing and binding of amyloid peptides, for joint therapy and diagnosis of Alzheimer’s disease (AD). Firstly, we characterized two in vitro models of BBB, based on immortalized cell lines of human and rat origin, the hCMEC/D3 and RBE4 cells, respectively. We tested the transendothelial electrical resistance (TEER) and the endothelial permeability (PE) of small hydrophilic compounds: our results, in agreement with data reported in literature, lead us to conclude that these cellular models are suitable for their employment as high-throughput screening tools. Subsequently, we tested NL mono-functionalized with three different peptides, the apolipoproteinE derived peptide (the ApoE monomer (mApoE), amino acids 141-150), its tandem dimer (dApoE) (141-150)2, and the Human Immunodeficency Virus type 1 (HIV-1) transactivator of transcription (TAT) peptide. We evaluated their uptake and PE; we selected the TAT functionalization as the best performing concerning cellular uptake, and the mApoE functionalization when considering both the internalization and PE. Once assessed the dynamics of mono-functionalized NL interactions with endothelial cells, we investigated mApoE- and dApoE-NL loading a curcumin-derivative (Re F. et al., 2011) to bind Aβ. We clearly demonstrated that the mApoE-functionalization allows a better drug cellular internalization, whereas dApoE-NL enhances drug PE at the highest extent. We then considered mApoE- and dApoE-NL exposing the Aβ targeting ligands phosphatidic acid (PA) or cardiolipin (CL), demonstrating that PA-mApoE-NL showed the highest cellular uptake and PE. We also studied TAT-NL exposing curcumin derivative3 (Airoldi C. et al., 2011) for Aβ binding, clearly indicating that TAT functionalization increased cellular uptake and PE of curcumin derivative3-NL. We also studied intracellular fate of NL double functionalized, exposing Aβ targeting ligands, and no co-localization was detected with acidic cellular compartments, suggesting that NL may escape from lysosomal degradative pathway. Taken together, these results indicate that the formulations herein analyzed are suitable tools for brain targeted drug and contrast agent delivery. We suggest further development of mApoE and dApoE-NL entrapped with a drug payload for their employment as BBB endothelial cell or brain targeted drug delivery tools, respectively. We also selected PAmApoE-NL and curcumin derivative3-TAT-NL as promising tools for their employment in combination for AD therapy and diagnosis. Further studies, based also on in vivo experiments, are needed to evaluate NL suitability for clinical exploitation. Finally, we inquired the endocytic mechanisms that mediates the entry of NL in the endothelial cells of BBB. We employed RNA interference technique to down-regulate caveolin1 expression. Our preliminary data suggest that caveolin1 and the related caveolaemediated endocytosis pathway may account for 40% of mApoE-NL cellular uptake. Future directions regard the down-regulation of other proteins specifically involved in different endocytic mechanisms, i.e. clathrin-mediated and adsoprptive endocytosis, in order to assess which endocytic mechanisms may account for ApoE and TAT-NL internalization.

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11

Silva, Juliana Galvão da. "Efeito dual de FGF2 e PMA em células HEK 293 transformadas por H-rasV12." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/46/46131/tde-20012015-085930/.

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Sabe-se há décadas que mutações nos genes ras estão presentes em cerca de 20% dos cânceres humanos, mas o desenvolvimento de terapias eficazes para o tratamento de câncer dependente dos oncogenes ras permanece um desafio científico importante. Nesse contexto, o nosso grupo publicou recentemente resultados interessantes mostrando que FGF2 exógeno ou PMA, contrariamente à expectativa geral, inibem a proliferação de células de camundongo malignas dependentes dos oncogenes H- ou K-Ras. Para dar continuidade a estes estudos o projeto desta tese foi planejado para investigar os mecanismos subjacentes a possíveis efeitos citotóxicos de FGF2 e PMA em células humanas transformadas por ras. Para esse fim, a linhagem humana imortalizada HEK 293 foi condicionalmente transformada pela expressão ectópica da construção quimérica de DNA ER:H-rasV12, que codifica a oncoproteína de fusão ER:H-RasV12, cuja atividade é induzível por 4-hidroxi-tamoxifen (4OHT). Essa abordagem nos permitiu verificar os efeitos de FGF2 e PMA em sublinhagens HEK/ER:HrasV12 fenotipicamente \"normais\" ou transformadas por níveis crescentes da oncoproteína H-RasV12. Os principais resultados mostraram que tanto FGF2 como PMA tem efeito dual promovendo ou inibindo a proliferação das células transformadas em função da concentração intracelular crescente de H-RasV12. Ensaios de crescimento de colônias em suspensão de agarose mostraram que: a) as células parentais HEK293 não desenvolveram colônias mesmo quando tratadas com FGF2 ou PMA, resultados que estão de acordo com seu fenótipo não tumoral; b) mas, as sublinhagens HEK/ER:HrasV12 deram origem a colônias mesmo quando tratadas com concentrações pequenas de 4OHT, que condicionaram níveis intracelulares baixos de ER:HRasV12; nestas condições experimentais, FGF2 foi um forte promotor do crescimento de colônias, condizente com sua reconhecida atividade promotora do crescimento de células tumorais em suspensão; ainda nestas condições, PMA não teve efeito significante sobre o crescimento de colônias; c) coerentemente, concentrações elevadas de 4-OHT levaram aos níveis intracelulares mais altos de ER:HRasV12 e, por conseguinte, a desenvolvimento máximo de colônias de células HEK/ER:HrasV12, no entanto, nestas condições, ambos FGF2 e PMA inibiram completamente o crescimento de colônias. Por outro lado, transformação de HEK293 com um vetor de expressão constitutiva de HrasV12 levou à seleção e isolamento das sublinhagens tumorais HEK/HrasV12, cujo fenótipo se caracterizou por: a) nenhum efeito de FGF2 sobre a sua proliferação e b) forte inibição de sua proliferação por PMA. A ação citotóxica de PMA exclusivamente observada em células HEK 293 transformadas por H-rasV12 se caracterizou por: a) total dependência de PKC, provavelmente mediada pela ativação proteolítica específica de PKC δ; b) envolvimento de níveis elevados e sustentados de ROS com disparo tardio de apoptose.
It is known for nearly 20 years that mutated ras oncogenes are found in 20% of human malignancies, however efficacious therapies are not yet available for Ras-driven cancer. Along of these lines, our group recently published provocative results showing, against common belief, that FGF2 and PMA inhibited proliferation of Ras-dependent malignant mouse cells. Aiming to gain insight into this intriguing phenomenon, the present thesis project was planned to investigate the possible cytotoxicity of FGF2 and PMA in human Ras-driven malignant cells. To this end an immortalized non-tumorigenic human cell line (HEK293) was stably transformed with the DNA construction ER:H-rasV12, which encodes the fusion protein ER:H-RasV12, whose activity requires activation by 4-hidroxitamoxifen (4-OHT). This approach allowed us to evaluate FGF2 and PMA effects on HEK/ER:HrasV12 sublines under switching from \"normal\" to transformed phenotypes upon 4-OHT induction. Our main results have shown that both FGF2 and PMA displayed dual effects promoting or inhibiting proliferation of HEK/ER:HrasV12 cells in function of ER:HRasV12 intracellular levels. Clonogenic assays in agarose suspension have shown: a) parental HEK293 line did not develop colonies under FGF2 and PMA treatment or not, in agreement with its non-tumorigenic nature; b) however, HEK/ER:HrasV12 sublines developed colonies even under low 4-OHT concentrations, which led to low ER:HRasV12 intracellular levels; under these conditions FGF2 strongly promoted colony growth and PMA had no effect; c) furthermore, in HEK/ER:HrasV12 sublines, elevated 4-OHT concentrations led to high ER:HRasV12 intracellular levels and maximal colony growth; but, under these experimental conditions both FGF2 and PMA abolished colony growth. On the other hand, HEK293 transformation with a vector that constitutively express HrasV12 yielded HEK/ER:HrasV12 sublines displaying the following phenotypic traits: a) non FGF2 effects on proliferation and b) severe proliferation inhibition by PMA. PMA toxicity, exclusively observed in HrasV12 -transformed HEK293 cells, was characterized by: a) total dependency on PKC, likely mediated by specific proteolytic activation of PKCδ; b) involvement of high and sustained ROS levels correlated with late apoptosis triggering.

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12

Tse, Wan-wai. "A study of genomic DNA methylation in immortalized human epithelial cell lines." Click to view the E-thesis via HKUTO, 2008. http://sunzi.lib.hku.hk/hkuto/record/B41290392.

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13

Tse, Wan-wai, and 謝韻慧. "A study of genomic DNA methylation in immortalized human epithelial cell lines." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2008. http://hub.hku.hk/bib/B41290392.

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14

Hasan, Shazeen Mumtaz. "Characterisation of immortalised human foetal retinal progenitor cell lines." Thesis, University College London (University of London), 2008. http://discovery.ucl.ac.uk/1444236/.

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Inherited retinal dystrophies and age-related macular degeneration (AMD) are leading causes of blindness. Current treatments only slow disease progression in a minority of patients, therefore it is important to develop new treatments that can regenerate lost cells, restore retinal function and halt disease progression. Retinal progenitor cells have the potential to replace degenerating photoreceptors in retinal diseases. This thesis explores the viability of using immortalised human foetal retinal progenitor cells for this purpose, and aims to elucidate the trophic factors required, in vitro, to drive these cells towards retinal cell lineages. We also examine their potential to survive, integrate and differentiate in the diseased and developing rat retina. Two human foetal retinal progenitor cell lines were established by infecting retinal foetal tissue with the temperature-sensitive tsT-SV40 antigen. The immortalised progenitor cells were compared to primary human foetal retinal progenitor cultures, and both were shown to express similar markers of neural progenitor cells and early neuronal cell types. Several trophic factors were investigated, including serum, retinoic acid and conditioned medium, with respect to their ability to influence gene expression. Serum appeared to induce expression of ganglion cell markers, and conditioned medium stimulated cell proliferation. Cells were also grafted into neonatal hooded Lister rats and RCS dystrophic rats, and neither the primary nor the immortalised progenitor cells demonstrated integration into the retina. In the RCS rat, cells were engulfed by host macrophage/microglia and showed no signs of integration or expression of neuronal markers. Immortalised cells, in vivo, stained positively for Ki67 and low levels of nestin, but exhibited limited ability to differentiate or integrate. These data indicate that immortalised progenitor cells maintain many characterisics of unimmortalised progenitor cells, and suggest that immortalisation of retinal progenitors may have long term therapeutic possibilities.

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15

Deurholt, Tanja. "Development of an immortalised human hepatocyte cell line for the AMC Bio-Artificial Liver." [S.l. : Amsterdam : s.n.] ; Universiteit van Amsterdam [Host], 2007. http://dare.uva.nl/document/54903.

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16

Williams, Meryl Marjory. "Transfection of human osteoblasts with SV40 DNA : characterisation of immortalised cell lines." Thesis, University of Bath, 1992. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.305405.

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17

Poppe, Sophia. "Establishment and investigation of tendon-derived cell lines immortalized by the human telomerase reverse transcriptase gene." Diss., lmu, 2010. http://nbn-resolving.de/urn:nbn:de:bvb:19-117327.

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18

Vasilyev, Vyacheslav V. "Regulation of gonadotropin [beta]-subunit gene expression by gonadotropin-releasing hormone in immortalized pituitary cell lines /." Diss., Connect to a 24 p. preview or request complete full text in PDF format. Access restricted to UC campuses, 2001. http://wwwlib.umi.com/cr/ucsd/fullcit?p3022216.

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19

Zhang, Zhiqi. "Blood-Brain Barrier in vitro Model: A Tissue Engineering Approach and Validation." FIU Digital Commons, 2010. http://digitalcommons.fiu.edu/etd/246.

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This dissertation evaluated the feasibility of using commercially available immortalized cell lines in building a tissue engineered in vitro blood-brain barrier (BBB) co-culture model for preliminary drug development studies. Mouse endothelial cell line and rat astrocyte cell lines purchased from American Type Culture Collections (ATCC) were the building blocks of the co-culture model. An astrocyte derived acellular extracellular matrix (aECM) was introduced in the co-culture model to provide a novel in vitro biomimetic basement membrane for the endothelial cells to form endothelial tight junctions. Trans-endothelial electrical resistance (TEER) and solute mass transport studies were engaged to quantitatively evaluate the tight junction formation on the in-vitro BBB models. Immuno-fluorescence microscopy and Western Blot analysis were used to qualitatively verify the in vitro expression of occludin, one of the earliest discovered tight junction proteins. Experimental data from a total of 12 experiments conclusively showed that the novel BBB in vitro co-culture model with the astrocyte derived aECM (CO+aECM) was promising in terms of establishing tight junction formation represented by TEER values, transport profiles and tight junction protein expression when compared with traditional co-culture (CO) model setups and endothelial cells cultured alone. Experimental data were also found to be comparable with several existing in vitro BBB models built from various methods. In vitro colorimetric sulforhodamine B (SRB) assay revealed that the co-cultured samples with aECM resulted in less cell loss on the basal sides of the insert membranes than that from traditional co-culture samples. The novel tissue engineering approach using immortalized cell lines with the addition of aECM was proven to be a relevant alternative to the traditional BBB in vitro modeling.

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Afsari, Farinaz. "The role of Wnt signalling pathway in mechanotransduction pathway in SV-40 immortalised human chondrocyte cell lines." Thesis, University of Edinburgh, 2005. http://hdl.handle.net/1842/28121.

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This thesis has set out to investigate whether Wnt pathway components are expressed in human chondrocyte cell lines and to explore whether the Wnt pathway is involved in mechanotransduction in chondrocytes. For the first time it is demonstrated that the Wnt signalling components, Wnt-1, Fz-2, Fzrp and β catenin are expressed in human chondocyte cell lines. Fibronectin and CD44 are identified in association with chondrocyte Wnt-Fz complexes suggesting that they may be coreceptors necessary for transducing Wnt signals intracellularly. The formation of GSK3β/β catenin degradation complexes was induced by a Wnt agonist and delayed by mechanical stimulation of chondrocytes, while, induction of GSK3β activity by mechanical stimulation was inhibited by a PI3K inhibitor. These experiments suggest that the Wnt signalling pathway may be involved in mechanical signalling in these cells. However, the induction of the GSK3β activity following mechanical stimulation is mediated by a PI3K dependent pathway rather than the Wnt pathway. The activity of GSK3β and the formation of GSK3β/β catenin complexes in chondrocytes are influenced by an interaction between mechanotransduction and the Wnt signalling. These, in turn, control the cytoplasmic/nuclear distribution of β-catenin which affects the regulation of downstream target genes such ad CD44, fibronectin and some metalloproteinases. CD44 and fibronectin are essential components of cartilage, which are involved in matrix assembly and the maintenance of cartilage integrity.

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21

Willett, Brian James. "The generation of immortalised cell lines from adult liver which retain the differentiated characteristics of the hepatocyte." Thesis, University of Strathclyde, 1989. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.280450.

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22

Hotchin, Neil Anthony. "A study of the effects of constitutive c-Myc expression in Epstein-Barr virus immortalised B cell lines." Thesis, Imperial College London, 1991. http://hdl.handle.net/10044/1/46825.

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23

Matloka, Magdalena. "MBNL derivatives for therapeutic application in myotonic dystrophy." Electronic Thesis or Diss., Sorbonne université, 2019. https://accesdistant.sorbonne-universite.fr/login?url=https://theses-intra.sorbonne-universite.fr/2019SORUS269.pdf.

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Les Dystrophies Myotoniques de type 1 (DM1) et 2 (DM2) sont des maladies neuromusculaires autosomiques dominantes causées par l’expansion anormale de séquences microsatellites C(C)TG situées respectivement dans la région 3’UTR du gène DMPK et le premier intron du gène ZNF9. Les ARNs mutés contenant les expansions de répétitions sont retenus dans le noyau des cellules sous formes d’agrégats riboprotéiques et séquestrent les protéines de liaison à l’ARN de la famille MBNL conduisant à des dérégulations de l’épissage alternatif associés aux symptômes cliniques. Bien que différentes approches thérapeutiques pour la DM soient en développement, il n’existe à ce jour aucune thérapie. Dans ce travail de thèse, nous avons développé une stratégie innovante de thérapie génique basée sur une protéine MBNL1 modifiée. Ce dérivé MBNL∆ agit comme un leurre pour libérer les facteurs MBNL endogènes anormalement séquestrés par les ARN mutés, et restaurer leurs fonctions. Ainsi l’expression de MBNL∆ dans des modèles DM1 permet la correction des anomalies moléculaires et phénotypiques. Nous avons également réalisé différentes optimisations de cet outil dans le but d’accroître ces capacités fonctionnelles. Enfin, nous avons développé un système d’autorégulation basé sur un « senseur d’épissage » dans le but de contrôler l’expression protéique d’un transgène. La preuve de concept de ce système a été faite à l’aide de MBNL∆ et validée dans des modèles DM1. En conclusion, mon travail de thèse a permis d’établir le potentiel d’une approche de thérapie génique pour la DM de type « leurre » basée sur l’ingénierie de protéine de liaison à l’ARN
Myotonic dystrophy (DM) is an autosomal neuromuscular disease encompassing two distinct forms, type 1 (DM1) and type 2 (DM2), which are caused by abnormal microsatellite expansions of C(C)TG repeats in the 3’UTR of the DMPK and first intron of ZNF9 genes, respectively. Mutant RNAs carrying expanded repeats are retained in the nucleus as riboprotein aggregates that abnormally sequester MBNL splicing factors leading to alternative splicing misregulations associated with clinical symptoms. Although various therapeutic approaches for DM are under development, there is no effective therapy available so far. In this study, we designed a novel gene therapy strategy with the use of an engineered MBNL RNA-binding protein derivative that acts as a CUGexp-decoy to release sequestered endogenous MBNL factors and restore their proper functions. Expression of the decoy results in the correction of DM1-associated features in both in vitro and in vivo models of the disease. Subsequent optimization processes were applied to the engineered decoy and the most potent derivate that increases its functional capacity was selected for further therapeutic application. Additionally, we developed an autoregulatory system based on a splice-sensor strategy to control transgene product expression and provided a proof-of-concept of its efficacy in both in vitro and in vivo systems. In conclusion, my work establishes the potency of gene therapy treatment for DM and support the use of the decoy-based approach as an alternate or complementary therapeutic intervention for DM

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24

Macnair, R. "A biocompatibility study of orthopaedic materials with primary and immortalised osteoblast-like cells derived from rat and human tissue." Thesis, University of Strathclyde, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.266231.

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25

Pardalejo, Mafalda Costa. "Differential Expression Profiling in an Immortalized Human Granulosa Cell Line (GC1a)." Master's thesis, 2018. https://hdl.handle.net/10216/118671.

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Pardalejo, Mafalda Costa. "Differential Expression Profiling in an Immortalized Human Granulosa Cell Line (GC1a)." Dissertação, 2018. https://hdl.handle.net/10216/118671.

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27

LI, RUI-NIAN, and 李瑞年. "Biochemical and biological characterization of a retrovirus isolated from an immortalized cell line." Thesis, 1989. http://ndltd.ncl.edu.tw/handle/00006815027110133869.

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JIANG, LIAN-CAI, and 蔣連財. "The establishment of a new human immortalized fibroblast cell line and its applications in medical research." Thesis, 1992. http://ndltd.ncl.edu.tw/handle/37533150521708960446.

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Chia-Wei and 陳嘉偉. "In vitro effects of citric acid on the induction of cell cycle arrest and apoptosis in human immortalized keratonocyte cell line (HaCaT)." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/07001396503784459387.

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碩士
中山醫學大學
醫學研究所
98
Cosmetic and skin care products have been hot topics in dermatology for decades. Alpha-hydroxyacids (AHAs) have been widely used. However, there is concern about its safety for long-term use. We had evaluated the potential cytotoxic effects of glycolic acid and lactic acid in previous studies. In this study, we investigated the cytotoxic effect of citric acid in human keratinocyte cell line (HaCaT). The study results suggested that citric acid was cytotoxic to HaCaT cells via the mechanisms of induction of apoptosis and cell cycle arrest. The results indicated that citric acid not only inhibited proliferation of HaCaT cells in a dose-dependent mode, but also induced apoptosis and cell cycle arrest at G2/M phase (before 24 hours) and S phase (at 48 hours). Western blotting results before 24 hours showed that citric acid decreases the levels of cyclin B, cdc2, cdc25c, P21 and increase the levels of chk2 that causes cell cycle arrest at G2/M phase. Western blotting results at 48 hours showed that citric acid increases the levels of cyclin A, cyclin E, CDK2, P21, P27 that cause cell cycle arrest at S phase. We observed cell morphological changes, and also used DAPI stain to evaluate DNA damage. The results of flow cytometry showed that citric acid induces HaCaT cell apoptosis through mitochondrial membrane potential (MMP) decrease and AIF, Endo G, and cytochrome c release from mitochondria to the cytosol. In addition, citric acid increased the level of Bax and inhibited the levels of Bcl-2, Bcl-xL and activated caspase 9 and caspase 3, which subsequently induced apoptosis via caspase-dependent and caspase-independent pathway. We also found that the decrease of MMP and the increase of reactive oxygen species (ROS) may be related to the increase of cytoplasmic calcium level. Citric acid also activated death receptor and increased the levels of caspase 8, or activated Bid protein. Therefore, citric acid could induce apoptosis through mitochondrial pathway and death receptor pathway in human kerationocyte cell line (HaCaT). Our study results suggested that the chemical exfoliation after applying citric acid might be caused by the induction of keratinocyte apoptosis. In the future, we will conduct further experiments to provide more academic basis for clinical application of hydroxyacids.

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Kuo, Shu Wen, and 郭書文. "Mechanism of Growth Restriction of Cell Line Immortalized by Temperature-Sensitive SV40 T Antigen at Non-Permissive Temperature." Thesis, 1995. http://ndltd.ncl.edu.tw/handle/17147499715765146519.

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碩士
國防醫學院
微生物及免役學研究所
83
When culcture temperature was shifted from 35 degrees celcius to 39 degrees celcius, human fibroblast immortalized by temperature sensitive SV40 T antigen became large and acquired morphological changes of senescent fibroblast. After culturing at 39 degrees for 48hrs, most cells cease to proliferation. A rapid increase of DNA content was observed after temperature shift. To elucidate the mechanism governing this rapid proliferation arrest, we studied the expression of genes involved in the regulation of cell cycle progression. Cyclin A and Cyclin B concentrations were not changed during growth restriction. Furthermore, we concluded that this growth arrest is not reversible. In these cells, p21 concertration remained high. This indicate that p21 protein was stabilized in growth resticted cells and suggest that p21 might be responsible for the mainternce of the use of growth restriction of immortalized fibroblast induced by temperature shift as a model system to study senescence.

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31

Greco, James. "The Effects of Fatty Acids on the Molecular Circadian Clock in Immortalized, Clonal Hypothalamic Neurons." Thesis, 2013. http://hdl.handle.net/1807/65436.

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Diets high in saturated fatty acids are associated with the development of circadian dysregulation, obesity, and type 2 diabetes mellitus. Conversely, unsaturated fatty acids are now known to improve insulin sensitivity, reduce weight gain, and alleviate obesity-induced inflammation. The aforementioned effects of saturated and unsaturated fatty acids have also been identified in the hypothalamus; however, there is a paucity of studies regarding the role of unsaturated fatty acids in circadian rhythms. Therefore, a novel cell model was established to examine the effects of omega-3 fatty acids on circadian rhythms in hypothalamic neurons. The mHypoE-37 cell line expresses Bmal1, Per2, and Rev-erbα in a circadian manner. The saturated fatty acid, palmitate, was found to induce circadian dysregulation of the mHypoE-37 neurons, whereas the unsaturated fatty acid, docosahexaenoic acid, protected against palmitate-induced circadian changes. These studies are the first to identify the potential for unsaturated fatty acids to protect the circadian system.

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Huang, Yu-Wei, and 黃昱瑋. "Study on the effects of curcumin and its analogs on the protection of hydrogen peroxide-treated human immortalized keratinocyte cell line (HaCaT)." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/gbbegy.

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碩士
臺北醫學大學
生藥學研究所
102
Hydrogen peroxide can produce large amounts of intracellular reactive oxygen species (reactive oxygen species, ROS) which can cause oxidative stress in cells. Many diseases and skin aging are associated with oxidative stress in cells. To find out which drug can reduce intracellular oxidative stress, hydrogen peroxide is used to treat cells for drug screenings. Curcumin, a polyphenol in turmeric, it has been reported to have various biological activities. In this study, we used curcumin and its analogs as materials to find out the protective activities against hydrogen peroxide-induced oxidative stress in HaCaT cells and the possible mechanisms. In the cell viability tests, it was found that the pretreatment of samples 7 and 24 at 5 μM for 2.5-h have significant protection against 150 μM hydrogen peroxide-induced apoptosis in human keratinocyte cell line (HaCaT cells). Using the flow cytometric analysis, it was found that the pretrement of samples 7 and 24 at 5 μM for 2.5-h could reduce the levels of early stage apoptosis in HaCaT cells. The pretreatment of samples 7 and 24 also reduce the expressions of p53 tumor suppressor protein, and reverse the loss of mitochondrial membrane potentials in HaCaT cells. The c-jun, c-fos, and iNOS mRNA expressions in HaCaT cells were also significantly decreased after being pretreated with samples 7 or 24. It was also found that the pretreatments of samples 7 and 24 at 5 μM for 2.5-h could significantly reduce intracellular ROS levels elevated by 150 μM hydrogen peroxide treatments. Using western blotting and enhanced chemiluminescence (ECL) system, it was found that the pretreatments of samples 7 and 24 could significantly induce heme oxygenase-1 (HO-1) expressions and glutathione peroxidase (GPx) activity in HaCaT cells. These results indicate that curcumin analogs, samples 7 and 24, show significantly protective effects against hydrogen peroxide-induced oxidative stress in HaCaT cells which may have protentials in cosmetics to develop anti-aging for skin cares.

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Gojska, Nicole. "Direct Steroidal Regulation and Inhibitory Mode of Action of Gonadotropin-inhibitory Hormone (GnIH or RFRP-3) in Immortalized Hypothalamic Cell Models." Thesis, 2013. http://hdl.handle.net/1807/65437.

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Fertility is dependent on the precisely orchestrated communication of an array of effectors within the reproductive axis, all of which impinge on gonadotropin-releasing hormone (GnRH) neurons. A novel reproductive inhibitor was identified in avian species and growing evidence suggests that the functional mammalian homologue, RFamide-related peptide-3 (RFRP-3 or GnIH) can inhibit GnRH neuronal activity and gonadotropin release. To date, the regulation and effects of RFRP-3 at the hypothalamic level are poorly understood. We established an Rfrp-expressing neuronal cell model to investigate the mechanisms of transcriptional regulation of the genes for RFRP and the RFRP receptor, GPR147 by dexamethasone and estradiol. We show that the RFRP system is a direct target for stress-associated transcriptional regulation. Further, employing a novel GnRH-secreting cell line, we report that GnRH neurons express Gpr147 and RFRP-3 represses the transcription of GnRH. These data further our understanding of the level and regulatory effects at which RFRP-3 modulates reproduction.

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34

Rekus, Martin T. [Verfasser]. "Characterization of growth and differentiation of a spontaneously immortalized keratinocyte cell line (HaCaT) in a defined, serum free culture system / vorgelegt von Martin T. Rekus." 2000. http://d-nb.info/963505262/34.

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35

"Molecular characterization of immortalized human uterine leiomyoma cell lines: A fibroid model." Tulane University, 2006.

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Human uterine leiomyomas (also known as fibroids, or myomas) are the most common benign tumors in women of reproductive age, affecting approximately 33% of all women and an estimated 77% of African American women. In the United States approximately 600,000 hysterectomies are performed annually at a cost of 1.7 billion dollars a year with fibroids accounting for 40% of procedures performed. Despite the high frequency of uterine leiomyomas and their major socioeconomic impact little research has been done in this area Many questions still remain to be answered concerning this disease. Therefore, we investigated a unique cell model to study uterine leiomyoma. First we characterized a novel immortalized cell line focusing on hormones and gene interactions. Cells were found to be estrogen responsive, and inhibited by ICI. High mobility group proteins are found in hormone responsive cancers such as breast, prostate, and benign uterine leiomyoma. We investigated an association between high mobility group proteins, estrogen, and estrogen receptors. From our data we concluded that high mobility group proteins are regulated by estrogen through the estrogen receptor pathway. Lastly using microarray technology we explored genes that may play a role in uterine leiomyoma growth. Our data suggest that growth of fibroids is not only activated through classic genomic pathways but also non-genomic pathways Our studies have tested a novel uterine leiomyoma cell line in hopes of establishing a universal model the field can use to study this important disease. Comparing our results with others in the field, we conclude that this cell line is an appropriate model for researching fibroids. Allowing researchers to explore molecular mechanisms involved in the etiology, growth and programmed cell death of these tumors
acase@tulane.edu

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36

Kim, Ginah. "The Hormonal Regulation of Kisspeptin and Neuropeptide Y Hypothalamic Neurons." Thesis, 2010. http://hdl.handle.net/1807/25727.

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Kisspeptin (encoded by Kiss1) is a hypothalamic neuropeptide that is directly regulated by sex steroids and directly stimulates gonadotropin-releasing hormone (GnRH) neurons. Kisspeptin cell models were established in order to facilitate future molecular analysis of kisspeptin. mHypoA-51 and mHypoA-63 cell lines were found to express kisspeptin, estrogen receptor α and β, substance P, but not tyrosine hydroxyase. Furthermore, estrogen decreased Kiss1 expression in both cell lines. Based on these results, it was concluded that mHypoA-51 and mHypoA-63 are representative of arcuate kisspeptin neurons. Accumulating evidence also indicates that kisspeptin indirectly stimulates GnRH neurons through afferent neurons. Kisspeptin receptor expression was detected in native neuropeptide Y (NPY) neurons. Using the mHypoE-38 cell line, kisspeptin was found to directly regulate NPY mRNA expression and secretion via the ERK1/2 and p38 MAPK pathways. This is the first evidence that kisspeptin directly stimulates NPY neurons to potentially exert indirect effects on GnRH neurons.

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37

Wachner, Sophia [Verfasser]. "Establishment and investigation of tendon-derived cell lines immortalized by the human telomerase reverse transcriptase gene / vorgelegt von Sophia Amina Poppe." 2010. http://nbn-resolving.de/urn:nbn:de:bvb:19-117327.

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Poppe, Sophia [Verfasser]. "Establishment and investigation of tendon-derived cell lines immortalized by the human telomerase reverse transcriptase gene / vorgelegt von Sophia Amina Poppe." 2010. http://d-nb.info/1005350159/34.

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39

Tung, Stephanie S. Y. "Serotonergic Responsiveness in Hypothalamic Neurons." Thesis, 2012. http://hdl.handle.net/1807/33771.

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Serotonin (5-HT) has been implicated in energy homeostasis. There is growing evidence that 5-HT, acting through the 5-HT1BR in the paraventricular nucleus of the hypothalamus (PVN), is important to this regulation. To investigate the cellular events underlying 5-HT1BR action, a PVN neuronal cell model was established. The mHypoA-2/30 cell line expresses a complement of markers and neuropeptides specifically localized to the PVN. 5-HT induces neuronal activation in a dose-dependent manner as determined by an elevation in cFos mRNA levels. As 5-HT exerted limited transcriptional control, the integrity of 5-HT signaling machinery was assessed. 5-HT signals through cAMP and calcium secondary messenger systems by suppressing cAMP and elevating intracellular calcium, effects that are mimicked by activating the 5-HT1BR and that are attenuated in the presence of inhibitors. These findings support the use of this novel PVN cell model for delineating components involved in direct 5-HT action in PVN neurons.

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